Equine faecal microbiota transplant: Current knowledge,proposed guidelines and future directions

While certainly not a novel concept, faecal microbiota transplant (FMT) has recently garnered renewed interest in veterinary medicine due to its remarkable success in treating recurrent Clostridium difficile infection (CDI) in man. There is a dearth of information on indications and efficacy of FMT for the treatment of gastrointestinal disorders in the horse; however, based on evidence in man and other veterinary species, and anecdotal reports in horses, FMT may be a useful treatment for selected cases of acute and chronic diarrhoea and inflammatory bowel disease (IBD) in the horse. In the absence of evidence, expert opinion is offered on case selection and FMT procedure. More research is needed to explore the efficacy, indications and optimal preparation, storage and delivery of FMT to horses.     

Preliminary molecular analysis of Clostridium difficile isolates from healthy horses in northern Italy

Clostridium difficile, associated with a wide spectrum of diseases in humans, as well as in several animal species, is an important cause of colitis in adult horses and foals. The aim of this study was to investigate by toxin gene profile and PCR-ribotyping the molecular characteristics of 14 C. difficile strains isolated from 42 faeces of healthy horses. Both toxin genes, tcdA and tcdB, were present in only 1 isolate (7.1%). Six isolates (42.9%) demonstrated tcdA−/tcdB+ genotype, and seven isolates (50.0%) were tcdA−/tcdB−. All strains were binary toxin genes negative (cdtA−/cdtB−). The PCR-positive strains, except for the tcdA+/tcdB+ isolate, tested negative for, in vitro, A and/or B toxins production by EIA. Eleven distinct ribotypes were observed.In conclusion, C. difficile can be present in the normal intestinal flora of healthy adult horses, in addition to foals. These animals could therefore play an important role as potential reservoirs of toxigenic strains.     

Equine hyperimmune serum protects mice against Clostridium difficile spore challenge

Clostridium (C.) difficile is a common cause of nosocomial diarrhea in horses. Vancomycin and metronidazole have been used as standard treatments but are only moderately effective, which highlights the need for a novel alternative therapy. In the current study, we prepared antiserum of equine origin against both C. difficile toxins A and B as well as whole-cell bacteria. The toxin-neutralizing activities of the antibodies were evaluated in vitro and the prophylactic effects of in vivo passive immunotherapy were demonstrated using a conventional mouse model. The data demonstrated that immunized horses generated antibodies against both toxins A and B that possessed toxin-neutralizing activity. Additionally, mice treated with the antiserum lost less weight without any sign of illness and regained weight back to a normal range more rapidly compared to the control group when challenged orally with 10⁷ C. difficile spores 1 day after serum injection. These results indicate that intravenous delivery of hyperimmune serum can protect animals from C. difficile challenge in a dose-dependent manner. Hence, immunotherapy may be a promising prophylactic strategy for preventing C. difficile infection in horses.     

Faecal shedding and serological cross‐sectional study of Lawsonia intracellularis in horses in the state of Minas Gerais,Brazil

Reason for performing the study: Proliferative enteropathy, caused by the intracellular bacterium Lawsonia intracellularis, has been described in horses in Australia, the USA, Canada and European countries but has not been reported in Latin America. The prevalence of the disease in horses worldwide is unknown. Objective: To evaluate the presence of subclinical L. intracellularis infection in horses in the state of Minas Gerais, Brazil. Methods: A longitudinal study using serology and PCR for detecting antibodies (IgG) and shedding of L. intracellularis in faecal samples, respectively, was conducted using a total of 223 horses from 14 different horse farms in Minas Gerais, and from the Veterinary School of UFMG equine herds in Minas Gerais. The immunoperoxidase technique in glass slides was used as the serological test. Results: Twenty‐one horse sera had immunoglobulin G titres of 1:60 and were considered positive. The PCR technique in faeces for L. intracellularis DNA identified 7 horses as faecal shedders. Horses shedding the organism appeared healthy, indicating that subclinical infection of L. intracellularis occurred in the horses. Conclusion: Seropositivity and detection of faecal shedding of L. intracellularis indicates the presence of the agent in the equine population in Minas Gerais. Potential relevance: Results of this study should alert clinicians in countries where proliferative enteropthy in horses has not been reported to consider this disease as a possible cause of enteric disease.     

Comparison of Clostridioides difficile strains from animals and humans: First results after introduction of C. difficile molecular typing and characterization at the Istituto Zooprofilattico Sperimentale of Piemonte,Liguria e Valle d’Aosta,Italy

PCR ribotypes (RTs027 and 078) are known causes of Clostridioides difficile infection (CDI) in humans. Molecular typing and characterization of 39 C. difficile strains isolated from samples from humas and animals in 2016–2018 indicated an overlap of RTs between community-acquired patients (CA-CDI) and domestic animals from the same geographical area; 14 RTs were identified: 12 RTs were positive for toxins A/B; RT078, RT080 and RT126 were also positive for binary toxin (CDT). Most of the RTs from the animals (RTs020, 078, 106, 126) were also detected in the samples from humans. Strains grouped into three clusters: cluster I included prevalently human strains, mainly RT 018; clusters II and III included strains from humans and animals, mainly RT078 and RT020. The CA-CDI strains suggested animals as a reservoir of C. difficile isolated together with other microorganisms from animals, highlighting the association of enteric pathogens as a cause of infection and death.     

Prevalence of Clostridium perfringens in faeces and ileal contents from grass sickness affected horses: Comparisons with 3 control populations

Reasons for performing study: While previous studies have demonstrated an association between equine grass sickness (EGS) and the presence of Clostridium botulinum within ileal contents and faeces, no such associations with other intestinal‐derived anaerobic bacteria have been extensively investigated. Hypothesis: The prevalence of C. perfringens in the ileal contents and faeces of EGS horses is greater than control horses; the detection of C. perfringens in faeces by ELISA could be diagnostically beneficial in a clinical setting. Methods: The prevalence of C. perfringens in faeces from EGS horses and healthy grazing control horses was determined by both selective culture and ELISA to permit both validation of the ELISA and inter‐group comparisons. Additionally, the prevalence of C. perfringens (ELISA) in ileal contents from EGS horses was compared with that for control horses with nongastrointestinal disease. Finally, the prevalence of C. perfringens (ELISA) in faeces from EGS cases was compared with that from both horses with which they shared pasture at the time of disease onset and non‐EGS colic horses. Results: When compared with culture, the ELISA had a sensitivity and specificity of 86 and 98%, respectively. The prevalence of C. perfringens in faeces as determined by both culture and ELISA was significantly higher (P<0.001) for EGS horses (7/9 and 15/37, respectively) than for healthy grazing controls (0/60 and 1/74, respectively). The prevalence of C. perfringens in ileal contents from EGS horses (5/10) was greater than that for horses with nongastrointestinal disease (1/12) at a level that approached significance (P = 0.056). EGS cases had a significantly greater prevalence of C. perfringens in faeces (15/37) than co‐grazing horses (1/18) and colic (1/16) horses. The specificity (93%) and PPV (94%) of the detection of C. perfringens by ELISA on faecal samples in relation to disease status (EGS compared with colic horses) was good. Sensitivity (41%) and NPV (39%) were poor. Conclusions and potential relevance: The use of a commercial ELISA to detect faecal C. perfringens may be diagnostically beneficial when differentiating EGS cases from colic cases, although further work is required to fully evaluate its potential.     

Typhlocolitis associated with <Emphasis Type="Italic">Clostridium difficile</Emphasis> ribotypes 078 and 110 in neonatal piglets from a commercial Irish pig herd

Background

Clostridium difficile is a recognised cause of typhlocolitis and diarrhoea in neonatal pigs but has never been confirmed in association with pathology and disease in Irish pigs.

Case Presentation

Four neonatal piglets, with a history of diarrhoea were referred to the Central Veterinary Research Laboratory, Backweston for necropsy. They were from a fully integrated, commercial pig farm with approximately 1000 sows. Three piglets had acute, superficial, erosive and suppurative typhlocolitis and the other had mild suppurative mesocolitis. Clostridium difficile (C. difficile) toxins A/B were detected using ELISA in the colonic contents from each piglet. C. difficile isolates from two of the piglets were PCR-ribotyped as 078 and an isolate from a third pig was ribotyped as 110.

Conclusions

This is the first report confirming C. difficile in association with typhlocolitis in Irish pigs.

     

Evaluation of five enzyme immunoassays compared with the cytotoxicity assay for diagnosis of Clostridium difficile-associated diarrhea in dogs.

Clostridium difficile-associated-diarrhea (CDAD) is a nosocomial infection in dogs. Diagnosis of this infection is dependent on clinical signs of disease supported by laboratory detection of C. difficile toxins A or B, or both, in fecal specimens via enzyme-linked immunosorbent assay (ELISA). Unfortunately, to the authors' knowledge, commercially available ELISAs have not been validated in dogs to date. We evaluated 5 ELISAs done on 143 canine fecal specimens (100 diarrheic and 43 nondiarrheic dogs) and on 29 C. difficile isolates. The results of each ELISA were compared with the cytotoxin B tissue culture assay (CTA). Clostridium difficile was isolated from 23% of the fecal specimens. Eighteen of the 143 fecal specimens were toxin positive (15 diarrheic and 3 nondiarrheic dogs). On the basis of multiplex polymerase chain reaction (PCR) analysis for toxin-A and -B genes, 72% of the isolates were toxigenic. The carriage rate of toxigenic isolates in diarrheic dogs was higher than that in the nondiarrheic dogs; however, these differences were not statistically significant. A good correlation was found between CTA, PCR, and culture results. The ELISAs done on fecal specimens collected from diarrheic dogs had low sensitivity (7-33%). In contrast, ELISA for toxin A or B, or both, performed on toxigenic isolates had high sensitivity (93%). These results suggest that commercially available human ELISAs are inadequate for the diagnosis of canine C. difficile-associated diarrhea when tested on fecal specimens. In contrast, the Premier ToxinA/B and Techlab ToxinA/B ELISAs may be useful for the diagnosis of canine CDAD when used on toxigenic isolates.     

Training young horses to social separation: effect of a companion horse on training efficiency

Reasons for performing study: The intensity with which a horse responds to separation from its group and subsequently to being alone is relevant for both horse and handler safety. Identification of training methods that may reduce responses to separation would be useful in practice. Objectives: To investigate whether the initial presence of a familiar companion horse modifies responses to separation from the group, lowers stress levels (as measured by heart rate) and increases training efficiency. Hypothesis: Habituation to separation proceeds more quickly if the horse is first trained with a companion, and heart rate is lower when the horse is subsequently trained alone, compared to control horses trained individually from the start. Methods: Young mares (n = 32), kept in groups of 4 were exposed to social separation: 2 horses of the group were trained singly (S1, n = 16) and the remaining 2 horses (n = 16) were trained first with a companion (P2) and then alone (P1). The training comprised 3 steps whereby distance from the group was gradually increased. The final learning criterion was met when a horse fed calmly alone inside a test arena (Step 3). Horses that were trained in a pair had to succeed in Step 3 together before they repeated the steps alone. Feeding behaviour and heart rate were recorded. Results: There were no significant differences between S1 and P1 horses, indicating that the initial pair‐training did not reduce responses in P1 horses. However, heart rate was significantly lower when horses were trained in pairs (P2) compared to when the same horses were subsequently trained alone (P1). Conclusions and potential relevance: It may not be efficient to habituate naïve young horses to social separation initially with a partner as these horses appear to have to relearn being in the test situation alone when switching to the individual training.     

Equine botulism: A clinical approach to diagnosis and management

Botulism is a syndrome of neuromuscular weakness caused by the toxins of Clostridium botulinum. Whilst it can affect most mammals, the horse appears to be one of the more susceptible species. Intoxication can occur via ingestion of preformed toxins in spoiled foodstuffs, ingestion of spores with colonisation in the intestinal tract or the contamination of wounds by C. botulinum. Food‐borne botulism is the most common worldwide, usually associated with spoiled roughage. Both individual cases and outbreaks have been reported, with generally a poor prognosis. Many affected horses succumb to recumbency and death/euthanasia shortly after onset of signs. Botulism should be considered a differential diagnosis for any horse displaying dysphagia or symmetrical neuromuscular weakness.     

An enzyme-linked immunosorbent assay (ELISA) test for the diagnosis of equine infectious anemia in Brazil

An enzyme-linked immunosorbent assay (ELISA) test was developed for the detection of specific antibodies against the unique infectious anemia (EIA) virus in equine sera. The ELISA test was faster and more sensitive when compared with the classic test of agar gel immunodiffusion (AGID). A total of 200 sera were tested: 100 from negative horses and 100 from positive horses by AGID. The ELISA test showed 92 horse sera negative and 100 horse sera positive by AGID with values of optical density (OD) less than 0.139 and higher than 0.139, respectively. Eight horse sera were negative by AGID and higher than 0.139 by ELISA. Six of these became AGID positive also when re-tested 30 days later, and two were of the horses that showed clinical signs of EIA and died before re-testing.     

Characterization of Clostridium perfringens in the feces of adult horses and foals with acute enterocolitis

Up to 60% of cases of equine colitis have no known cause. To improve understanding of the causes of acute colitis in horses, we hypothesized that Clostridium perfringens producing enterotoxin (CPE) and/or beta2 toxin (CPB2) are common and important causes of severe colitis in horses and/or that C. perfringens producing an as-yet-undescribed cytotoxin may also cause colitis in horses. Fecal samples from 55 horses (43 adults, 12 foals) with clinical evidence of colitis were evaluated by culture for the presence of Clostridium difficile, C. perfringens, and Salmonella. Feces were also examined by enzyme-linked immunosorbent assay (ELISA) for C. difficile A/B toxins and C. perfringens alpha toxin (CPA), beta2 toxin (CPB2), and enterotoxin (CPE). Five C. perfringens isolates per sample were genotyped for the following genes: cpa, cpb, cpb2 consensus, cpb2 atypical, cpe (enterotoxin), etx (epsilon toxin), itx (iota toxin), netB (necrotic enteritis toxin B), and tpeL (large C. perfringens cytotoxin). The supernatants of these isolates were also evaluated for toxicity for an equine cell line. All fecal samples were negative for Salmonella. Clostridium perfringens and C. difficile were isolated from 40% and 5.4% of samples, respectively. All fecal samples were negative for CPE. Clostridium perfringens CPA and CPB2 toxins were detected in 14.5% and 7.2% of fecal samples, respectively, all of which were culture-positive for C. perfringens. No isolates were cpe, etx, netB, or tpeL gene-positive. Atypical cpb2 and consensus cpb2 genes were identified in 15 (13.6%) and 4 (3.6%) of 110 isolates, respectively. All equine C. perfringens isolates showed far milder cytotoxicity effects than a CPB-producing positive control, although cpb2-positive isolates were slightly but significantly more cytotoxic than negative isolates. Based on this studied population, we were unable to confirm our hypothesis that CPE and CPB2-producing C. perfringens are common in horses with colitis in Ontario and we failed to identify cytotoxic activity in vitro in the type A isolates recovered.     

Serologic and molecular survey of horses to Coxiella burnetii in East of Iran a highly endemic area

There are few reports about Q fever in horse populations worldwide. This study aimed to detect the C. burnetii infection by serologic and molecular confirmation using commercial ELISA kit and real-time PCR in the East of Iran a region highly endemic. A total of 177 blood samples and 115 vaginal swabs were randomly collected from horses in East of Iran. The sera samples were analyzed for anti C.burnetii Ig G antibodies by a commercial ELISA kit and nucleic acid extraxted from vaginal samples were used to determine the C. burnetii DNA by real-time PCR assay. Antibodies were detected in 5.64 % (10/177) of sera samples and C. burnetii DNA was detected in 7.82 % (9/115) of horse vaginal samples. There was no significant difference in seroprevalence in different sex, age and breed groups. Our study showed that horses could be considered as a mild potential reservoir of C. burnetii which may be effective on horse health status. However, additional studies are needed to assess whether the horse could be considered as a relevant transmission risk indicator for Q fever.     

Bacterial probiotics as an aid in the control of Clostridium difficile disease in neonatal pigs

Although Clostridium difficile infection (CDI) is a common disease in swine, there is a lack of prevention strategies. The objectives of this study were to evaluate: i) the effectiveness of Lactobacillus spp. and ii) non-toxigenic C. difficile (NTCD) as prevention for the development of CDI in piglets. Cesarean-derived piglets (N = 150) were randomly assigned to 6 groups: GROUP 1 — negative control (n = 10); GROUP 2 — NTCD only (n = 13); GROUP 3 — Lactobacillus spp. only (n = 14); GROUP 4 — positive control (challenged with toxigenic C. difficile strain) (n = 35); GROUP 5 — NTCD and challenged with the toxigenic C. difficile strain (n = 34); and GROUP 6 — Lactobacillus spp. and challenged with the toxigenic C. difficile strain (n = 44). Piglets which received NTCD showed lower prevalence of toxin-positive feces, mesocolonic edema, and microscopic lesions compared with positive control piglets. Administration of Lactobacillus spp. did not reveal clear benefits.     

Re-emergence of Pigeon Fever (Corynebacterium pseudotuberculosis) Infection in Texas Horses: Epidemiologic Investigation of Laboratory-Diagnosed Cases

In recent years, Texas has seen a dramatic increase in the number of clinical cases of Corynebacterium pseudotuberculosis (pigeon fever) infection in horses. Equine pigeon fever cases at Texas Veterinary Medical Diagnostic Laboratories (TVMDL) were analyzed with the objectives of investigating the spatiotemporal distribution and seasonal and annual trends of pigeon fever infection in horses in Texas between 2005 and 2011 and identifying high-risk areas and create a risk map for pigeon fever in horses in Texas. The study population consisted of horses culture-positive for C. pseudotuberculosis between January 1, 2005, and December 31, 2011 at TVMDL. The Poisson model of scan statistics was fitted to identify disease clusters. Empirical Bayesian smoothing was performed with the crude incidence estimates, followed by the geostatistical method of kriging to delineate high-risk areas. Cases increased 10-fold between 2005 and 2011. The annual cumulative incidence ranged from 9.3 to 99.5 per 100,000 horses at risk. Two seasonal peaks in the number of cases were observed in June and in December. Scan statistics identified a primary cluster in central Texas in 2011 (P < .0001 and relative risk of 9.2). Isopleth risk mapping also delineated a high-risk area in central Texas. High-risk areas were also detected in the panhandle and northern Texas. The epidemiological investigation supported anecdotal reports that pigeon fever is re-emerging in the Texas horse population. This study provides a baseline for future investigations of pigeon fever in the Texas horse population and serves as a reference for the disease distribution for veterinarians and horse owners.     

Crib‐biting in US horses: Breed predispositions and owner perceptions of aetiology

Reasons for performing study: Crib‐biting is an equine stereotypy that may result in diseases such as colic. Certain breeds and management factors have been associated. Objectives: To determine: breed prevalence of crib‐biting in US horses; the likelihood that one horse learns to crib‐bite from another; and owner perceptions of causal factors. Methods: An initial postal survey queried the number and breed of crib‐biting horses and if a horse began after being exposed to a horse with this habit. In a follow‐up survey, a volunteer subset of owners was asked the number of affected and nonaffected horses of each breed and the extent of conspecific contact. The likelihood of crib‐biting given breed and extent of contact was quantified using odds ratio (OR) and significance of the association was assessed using the Chi‐squared test. Results: Overall prevalence was 4.4%. Thoroughbreds were the breed most affected (13.3%). Approximately half of owners believed environmental factors predominantly cause the condition (54.4%) and crib‐biting is learned by observation (48.8%). However, only 1.0% of horses became affected after being exposed to a crib‐biter. The majority (86%) of horses was turned out in the same pasture with other horses and extent of contact with conspecifics was not statistically related to risk. Conclusion: This is the first study to report breed prevalence for crib‐biting in US horses. Thoroughbreds were the breed more likely to be affected. More owners believed either environmental conditions were a predominant cause or a combination of genetic and environmental factors contributes to the behaviour. Only a small number of horses reportedly began to crib‐bite after being exposed to an affected individual, but approximately half of owners considered it to be a learned behaviour; most owners did not isolate affected horses. Potential relevance: Genetic predisposition, not just intensive management conditions and surroundings, may be a factor in the high crib‐biting prevalence in some breeds, and warrants further investigation. Little evidence exists to suggest horses learn the behaviour from other horses, and isolation may cause unnecessary stress.     

Prevalence of anti-<Emphasis Type="Italic">Toxoplasma gondii</Emphasis> antibodies in sport horses from Qazvin,Iran

In the present study, the prevalence of antibodies to Toxoplasma gondii in sport horses of Qazvin was examined using modified agglutination test (MAT). On 52 horse sera totally examined for anti-Toxoplasma antibodies, 37 horses (71.2%) were seropositive by MAT. Results of the present study showed a high rate of Toxoplasma infection in horses in Qazvin area. More comprehensive study on equine toxoplasmosis is recommended.     

Quantitative and qualitative comparison of three scoring systems for assessing recovery quality after general anaesthesia in horses

Objective To assess the reproducibility and repeatability of two commonly used recovery quality scoring systems and compare them with those of a novel system based on a greater number of objective criteria. Animals The video‐recorded recoveries of ten client‐owned horses selected from all recovery recordings taken between September 2005 and March 2006 at the Royal (Dick) School of Veterinary Studies. Materials and methods A digital versatile disc (DVD) was produced using edited video recordings of ten horses recovering from general anaesthesia. Twelve experienced equine anaesthetists (raters) studied the DVD on three occasions, and scored the recovery quality of each horse using one of three scoring systems (P, D or E) on each occasion. The process was repeated 6 months later (t = 6) to measure intra‐observer reliability (repeatability). At first use (t = 0) raters were asked to comment on the advantages and disadvantages of each system. Results Inter‐rater variability was limited for each system: at each observation period raters accounted for 0.3–4.4% variation. System P was insensitive to differences between recoveries. In system D, score variability increased as recovery quality deteriorated. Intra‐rater variability varied with system: using system P, raters provided consistent scores between the observation periods for some, but not all horses (‘horse’ and ‘rater’ accounted for 9.7% and 1.9% of variation respectively). Raters were less consistent between t = 0 and t = 6 using system D, but each horse was scored with similar consistency. System E produced little variation at the level of horse (1.0%) and rater (1.9%). Raters broadly agreed on the principle advantages and disadvantages of the three systems. Conclusions and clinical relevance The systems examined showed reliability and reproducibility but practicality and simplicity of use appeared to be inextricably linked with imprecision. Further work is required to produce a suitable recovery quality scoring system.     

Injuries and outcome of three horses hospitalised in the aftermath of Hurricane Isaac

This case series describes 3 horses rescued from flooded areas, hospitalised and treated at the Louisiana State University Veterinary Teaching Hospital in the wake of Hurricane Isaac. The purpose of the paper is to familiarise practitioners with the injuries that may be found in flood‐afflicted horses and livestock. Case 1 presented with a degloving injury of the right hindlimb. The gelding developed neurological signs and was subjected to euthanasia; necropsy was consistent with Salmonella sepsis and disseminated mycosis due to Candida krusei, the first case reported in a horse. Case 2 presented with dermatitis attributable to chemical exposure and pneumonia and was subjected to euthanasia due to severe bilateral laminitis. Case 3 sustained a degloving injury of the left hindlimb and was subjected to euthanasia due to intractable pain. This case series illustrates the need for preparation and evacuation to avoid life‐threatening injuries in equids and livestock caused by hurricanes and their aftermath.